Determining factors for optimal neuronal and glial Golgi-Cox staining
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ORIGINAL PAPER
Determining factors for optimal neuronal and glial Golgi‑Cox staining Sareesh Naduvil Narayanan1 · Laxminarayana Kurady Bairy2 · Suresh Kumar Srinivasamurthy2 Accepted: 30 May 2020 © Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Golgi staining allows for the analysis of neuronal arborisations and connections and is considered a powerful tool in basic and clinical neuroscience. The fundamental rules for improving neuronal staining using the Golgi-Cox method are not fully understood; both intrinsic and extrinsic factors may control the staining process. Therefore, various conditions were tested to improve the Golgi-Cox protocol for vibratome-cut rat brain sections. Optimal staining of cortical neurons was achieved after 72 h of impregnation. Well-stained neurons in both cortical and subcortical structures were observed after 96 h of impregnation. The dendritic arborisation pattern of cortical neurons derived from the 72-h impregnation group was comparable to those of the 96 and 168-h impregnation groups. The entire brain was stained well when the pH of the Golgi-Cox solution was 6.5 and that of the sodium carbonate solution was 11.2. Lack of brain perfusion or perfusion with 0.9% NaCl did not influence optimal neuronal staining. Perfusion with 37% formaldehyde, followed by impregnation, only resulted in glial staining, but perfusion with 4% formaldehyde facilitated both glial and neuronal staining. Whole brains required longer impregnation times for better staining. Although every factor had a role in determining optimal neuronal staining, impregnation time and the pH of staining solutions were key factors among them. This modified Golgi-Cox protocol provides a simple and economical procedure to stain both neurons and glia separately. Keywords Golgi-Cox staining · Neuron · Glia · Cortex · Hippocampus · Formalin
Introduction Golgi’s (1873) ‘black reaction’ histological staining method was the first to allow a full view of a neuron’s structure. More than 146 years later, it is still considered one of the best neurohistological staining techniques for determining the complete cytoarchitecture of neurons (Hinova-Palova et al. 2019; Vints et al. 2019; Zhang et al. 2003) and different brain regions. The fascinating feature of this brain staining technique is that it stains a random number of neurons (3–5%) by an unexplained mechanism, leaving the rest of the tissue unstained (Spacek 1989). This special feature of the Golgi staining technique enabled Santiago Ramón y Cajal to
* Sareesh Naduvil Narayanan [email protected]; [email protected] 1
Department of Physiology, RAK College of Medical Sciences, RAK Medical and Health Sciences University, PO Box 11172, Ras Al Khaimah, United Arab Emirates
Department of Pharmacology, RAK College of Medical Sciences, RAK Medical and Health Sciences University, PO Box 11172, Ras Al Khaimah, United Arab Emirates
2
study, and extensively describe, the cellular architecture of the vertebrate nervous system (Cajal 1909, 1910). Chang
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